C jun rabbit mab

Cells were washed once with PBS and then lysed in buffer containing 100 mM Tris-HCL (pH 6.8), 200 mmol/L DTT, 4% SDS, 20% glycerine. Protein concentrations in the supernatants were determined by the Bradford method. Equal amounts of protein from whole-cell lysates were separated by gel electrophoresis on 10% gels, transferred to PVDF membranes and were probed with specific primary antibody [MMP-1goat mAb; MMP-3 mouse mAb; Pi-ATF-2 mouse mAb, ATF-2 rabbit mAb (Santa Cruz, CA), Pi-Erk rabbit mAb, Erk rabbit mAb, Pi-Jnk rabbit mAb, Jnk rabbit mAb, Pi-P38 rabbit mAb, p38 rabbit mAb, Pi-c-Jun rabbit mAb, c-Jun rabbit mAb, C-Fos rabbit mAb, Pi-C-Fos rabbit mAb (Cell Signaling, MA) and then with the appropriate HRP-conjugated secondary antibodies. Proteins were detected using the enhanced chemiluminescence detection kit (Thermal Science, USA). For loading control, the membrane was probed with a monoclonal antibody for GAPDH (Kangchen Biotechnology, Shanghai). The quantitative analysis of the proteins was performed by normalizing against GAPDH or total protein by using Image Pro Plus 6.0 software (Media Cybernetics, Inc., Silver Spring, MD, USA).